p53 (Phospho-Ser315)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie designed for the detection of phosphorylated p53 at serine 315 in cultured cells. The assay uses a colorimetric readout at 450 nm and is reactive with human samples, with a stated dynamic range of > 5000 Cells. It supports measurement of the relative amounts of phosphorylated p53 in adherent cells and can be used to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on p53 phosphorylation.

Detection is achieved by an indirect ELISA format. Phospho-Ser315 p53 is captured by target-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Addition of substrate produces a colorimetric reaction that is read at 450 nm. The kit is described by the manufacturer as a convenient, lysate-free, high-throughput and sensitive assay for profiling p53 protein phosphorylation and expression in cells.

For data normalization, a monoclonal antibody specific for human GAPDH is included to serve as an internal positive control, and a Crystal Violet whole-cell staining step may be used to determine cell density so that absorbance values can be normalized to cell amounts. Phosphorylation of p53 at serine 315 has been implicated in DNA damage response, cell cycle regulation, and apoptosis, with dysregulation associated with diseases including cancer. The target is human TP53 (Gene ID: 7157, UniProt ID: P04637), and the sub-type of the assay is phospho-specific.